17β-hydroxysteroid dehydrogenase type 3 inhibitors for the treatment of androgen dependent diseases

ABSTRACT

In its many embodiments, the present invention provides a novel class of compounds as inhibitors of type 3 17β-hydroxysteroid dehydrogenase, methods of preparing such compounds, pharmaceutical compositions containing one or more such compounds, methods of preparing pharmaceutical formulations comprising one or more such compounds, and methods of treatment, prevention, inhibition, or amelioration of one or more diseases associated with type 3 17β-hydroxysteroid dehydrogenase using such compounds or pharmaceutical compositions.

FIELD OF THE INVENTION

This application is a divisional of U.S. application Ser. No.11/360,711, filed Feb. 23, 2006, which is a divisional of U.S.application Ser. No. 10/735,983, filed Dec. 15, 2003, and claims thebenefit of U.S. Provisional Application Ser. No. 60/434,101, filed Dec.17, 2002.

BACKGROUND OF THE INVENTION

Androgen dependent diseases, for example, diseases whose onset orprogress is aided by androgenic activity, are well known. These diseasesinclude, but are not limited to, prostate cancer, benign prostatichyperplasia, acne, seborrhea, hirsutism, androgenic alopecia, precociouspuberty, adrenal hyperlasia and polycystic ovarian syndrome. Estrogendependent diseases, for example, diseases whose onset or progress isaided by estrogenic activity, are also well known. These include, butare not limited to, breast cancer, endometriosis, leiomyoma andprecocious puberty.

Androgenic and estrogenic activities can be suppressed by administering,respectively, androgen and estrogen receptor antagonists. See, for e.g.,WO 94/26767 and WO 96/26201. Androgenic and estrogenic activities canalso be reduced by suppressing androgen and estrogen biosyntheses usinginhibitors of enzymes that catalyze one or more steps of suchbiosyntheses. 17β-HSD3 is the primary enzyme that convertsandrostenedione to testosterone in the testes. Inhibitors of both Type 3and Type 5 17β-hydroxysteroid dehydrogenase are described in WO99/46279. Inhibitors of Type 5 17β-hydroxysteroid dehydrogenase is alsodescribed in WO 97/11162. Androgenic and estrogenic activities can alsobe reduced by suppressing ovarian or testicular secretions by knownmethods. See, for e.g., WO 90/10462, WO 91/00731, WO 91/00733 and WO86/01105.

Commonly owned, pending U.S. patent applications Ser. No. 10/235,627,filed Sep. 5, 2002, and Ser. No. 10/271,358, filed Oct. 15, 2002,disclose certain types of inhibitors of type 3 17β-hydroxysteroiddehydrogenase. Pending provisional patent application Ser. No.60/427,363, filed Nov. 18, 2002, discloses certain types of inhibitorsof type 3 17β-hydroxysteroid dehydrogenase too.

There is a continuing need for new compounds, formulations, treatmentsand therapies to treat diseases and disorders associated with type 317β-hydroxysteroid dehydrogenase. It is, therefore, an object of thisinvention to provide compounds useful in the treatment or prevention oramelioration of such diseases and disorders.

SUMMARY OF THE INVENTION

In its many embodiments, the present invention provides a novel class ofcompounds as inhibitors of type 3 17β-hydroxysteroid dehydrogenase,methods of preparing such compounds, pharmaceutical compositionscomprising one or more such compounds, methods of preparingpharmaceutical compositions comprising one or more such compounds, andmethods of treatment, prevention, inhibition or amelioration of one ormore diseases associated with type 3 17β-hydroxysteroid dehydrogenaseusing such compounds or pharmaceutical compositions.

In one aspect, the present application discloses a compound, or apharmaceutically acceptable salt or solvate of said compound, saidcompound having the general structure shown in Formula I:

wherein:

-   X is CH or N;-   Y is selected from the group consisting of C, CH or N, and when Y is    CH or N, the optional covalent bond (represented by the dotted line    between rings marked II and IV) is absent, and when Y is C, that    optional covalent bond is present;-   G is (CHR⁴)_(n) or C(═O);-   R is selected from the group consisting of alkyl, —OR⁴, aryl,    heteroaryl, heteroarloxy, heterocyclyl, heterocyclyloxy, cycloalkyl,    cycloalklyloxy, —N(R⁴)₂ where the two R⁴ moieties can be the same or    different, —(CH₂)_(n)-aryl, —(CH₂)_(n)-heteroaryl,    —(CH₂)_(n)-heterocyclyl and —(CH₂)_(n)-cycloalkyl, wherein each of    said alkyl, aryl, heteroaryl, heterocyclyl and cycloalkyl can be    unsubstituted or optionally independently substituted with one or    more moieties which can be the same or different, each being    independently selected from the group consisting of alkyl, aryl,    heteroaryl, —OR⁴, heterocyclyl, heterocyclyloxy, cycloalkyl,    cycloalklyloxy, —N(R⁴)₂ where the two R⁴ moieties can be the same or    different, —C(O)R⁴, and —C(O)N(R⁴)₂ where the two R⁴ moieties can be    the same or different;-   one of a, b, c and d represents N or N⁺O⁻, and the remaining a, b, c    and d groups represent C(R¹) or C(R²); or    -   each of a, b, c, and d are independently selected from C(R¹) or        C(R²);-   R¹ and R² can be the same or different, each being independently    selected from the group consisting of:    -   H, halo, —CF₃, —OR⁴, —C(O)R⁴, —OCF³, —SR⁴, —S(O)_(n)R⁵,        benzotriazol-1-yloxy, tetrazol-5-ylthio, alkynyl, alkenyl        wherein said alkenyl can be unsubstituted or optionally        substituted with halo, —OR⁴ or —C(O)OR⁴, alkyl wherein said        alkyl can be unsubstituted or optionally substituted with halo,        —OR⁴ or —C(O)OR⁴, —N(R⁴)₂ where the two R⁴ moieties can be the        same or different, —NO₂, —OC(O)R⁵, —C(O)OR⁴, —CN, —N(R⁴)C(O)OR⁴,        —SR⁵C(O)OR⁴, and —SR⁵N(R⁴)₂ (provided that R⁵ in —SR⁵N(R⁴)₂ is        not —CH₂—) wherein each R⁴ is independently selected;-   the dotted line between carbon atoms 5 and 6 represents an optional    bond, such that when a double bond between carbon atoms 5 and 6 is    present, A and B can be the same or different, each being    independently selected from the group consisting of —R⁴, halo, —OR⁴,    —C(O)OR⁴, —OC(O)OR⁴ or —OC(O)R⁴, and when no double bond is present    between carbon atoms 5 and 6, A and B can be the same or different,    each being independently selected from the group consisting of (H₂),    —(OR⁵)₂, (H and halo), (dihalo), (H and R⁵), (R⁵)₂, (H and    —OC(O)R⁴), (H and —OR⁴), (═O), and (H, (═NOR⁴) or (—O—(CH₂)_(p)—O—)    wherein p is 2, 3 or 4);-   R³ is selected from the group consisting of H, alkyl, alkoxy and    alkoxyalkyl;-   R⁴ is selected from the group consisting of H, alkyl, aryl and    aralkyl;-   R⁵ is alkyl or aryl;-   R⁶ is H or alkyl;-   n is a number from 1-4; and-   q is a number from 1-8.

The compounds of Formula I can be useful as inhibitors of type 317β-hydroxysteroid dehydrogenase and can be useful in the treatment andprevention of diseases associated with type 3 17β-hydroxysteroiddehydrogenase.

DETAILED DESCRIPTION

In one embodiment, the present invention discloses compounds which arerepresented by structural Formula i, or a pharmaceutically acceptablesalt or solvate thereof, wherein the various moieties are as describedabove.

In a preferred embodiment, position a in ring I is N or N⁺O⁻,

In another preferred embodiment, A and B in ring II are H₂, i.e., theoptional bond is absent, and the C5-C6 bridge is unsubstituted.

In another preferred embodiment, R¹ and R² can be the same or different,each being independently H or halo.

In another preferred embodiment, R is selected from the group consistingof unsubstituted alkyl, alkyl substituted with a heterocyclyl, —N(R⁴)₂where the two R⁴ moieties can be the same or different, and —OR⁴,wherein said heterocyclyl can be unsubstituted or optionally substitutedwith one or more moieties which can be the same or different, each beingindependently selected from the group consisting of alkyl, aryl, —OR⁴,—N(R⁴)₂ where the two R⁴ moieties can be the same or different, —C(O)R,and —C(O)N(R⁴)₂ where the two R⁴ moieties can be the same or different.

In another preferred embodiment, X is CH.

In another preferred embodiment, X is N.

In another preferred embodiment, Y is N.

In another preferred embodiment, Y is CH.

In another preferred embodiment, R³ is H and q is 8.

In another preferred embodiment, R³ is alkyl and q is 1.

In another preferred embodiment, R³ is alkoxyalkyl and q is 1.

In another preferred embodiment, R³ is aralkyl.

In another preferred embodiment, R⁴ is H, alkyl or aryl.

In another preferred embodiment, R⁵ is alkyl.

In another preferred embodiment, R⁶ is H.

In an additional preferred embodiment, position a in ring I is N.

In an additional preferred embodiment, position a in ring I is N andpositions b, c and d are all the same and are C(R¹).

In an additional preferred embodiment, R¹ and R² are the same ordifferent, each being independently selected from H, Br, F and Cl.Non-limiting, illustrative substitutions on rings I and III of formula 1include trihalo, dihalo and monohalo substituted compounds, such as, forexample: (i) 3,8,10-trihalo; (ii) 3,7,8-trihalo; (iii) 3,8-dihalo; (iv)8-halo; (v) 10-halo; and (vi) 3-halo (i.e., no substituent in Ring III)substituted compounds; wherein each halo is independently selected.Preferred compounds of formula I include: (1)3-Br-8-Cl-10-Br-substituted compounds; (2) 3-Br-7-Br-8-Cl-substitutedcompounds; (3) 3-Br-8-Cl-substituted compounds; (4)3-Cl-8-Cl-substituted compounds; (5) 3-F-8-Cl-substituted compounds; (6)8-Cl-substituted compounds; (7) 10-Cl-substituted compounds; (8)3-Cl-substituted compounds; (9) 3-Br-substituted compounds; and (10)3-F-substituted compounds.

In an additional preferred embodiment, R is selected from the groupconsisting of unsubstituted alkyl, alkyl substituted with aheterocyclyl, —NH₂, and t-butoxy, wherein said heterocyclyl can beunsubstituted or optionally substituted with one or more moietiesselected from the group consisting of

-   —C(O)alkyl, and —C(O)N(alkyl)₂ where the two alkyl moieties can be    the same or different.

In an additional preferred embodiment, R³ is tert-butyl and q is 1.

In an additional preferred embodiment, R³ is 2-(methoxy)ethyl and q is1.

In an additional preferred embodiment, R³ is n-butyl and q is 1.

In an additional preferred embodiment, R³ is benzyl and q is 1.

In an additional preferred embodiment, R⁴ is H.

In an additional preferred embodiment, R⁴ is alkyl.

In an additional preferred embodiment, R⁵ is methyl.

In an additional preferred embodiment, n is 1.

In an additional preferred embodiment, q is 1.

In an additional preferred embodiment, p is 1.

A particularly preferred group of compounds are shown in Table 1.

TABLE 1

As used above, and throughout this disclosure, the following terms,unless otherwise indicated, shall be understood to have the followingmeanings:

“Patient” includes both human and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight orbranched and comprising about 1 to about 20 carbon atoms in the chain.Preferred alkyl groups contain about 1 to about 12 carbon atoms in thechain. More preferred alkyl groups contain about 1 to about 6 carbonatoms in the chain. Branched means that one or more lower alkyl groupssuch as methyl, ethyl or propyl, are attached to a linear alkyl chain.“Lower alkyl” means a group having about 1 to about 6 carbon atoms inthe chain which may be straight or branched.

“Aryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 6 to about 14 carbon atoms, preferably about 6 to about10 carbon atoms. Non-limiting examples of suitable aryl groups includephenyl and naphthyl.

“Heteroaryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 5 to about 14 ring atoms, preferably about 5 to about10 ring atoms, in which one or more of the ring atoms is an elementother than carbon, for example nitrogen, oxygen or sulfur, alone or incombination. Preferred heteroaryls contain about 5 to about 6 ring atomsThe prefix aza, oxa or thia before the heteroaryl root name means thatat least a nitrogen, oxygen or sulfur atom respectively, is present as aring atom A nitrogen atom of a heteroaryl can be optionally oxidized tothe corresponding N-oxide. Non-limiting examples of suitable heteroarylsinclude pyridyl, N-oxide of pyridyl, pyrazinyl, furanyl (furyl),thienyl, pyrimidinyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl,pyrazolyl, furazanyl, pyrrolyl, pyrazolyl, triazolyl,1,2,4-thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl,imidazo[1,5-a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl,indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl,imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl,pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl,1,2,4-triazinyl, benzothiazolyl and the like.

“Aralkyl” or “arylalkyl” means an aryl-alkyl- group in which the aryland alkyl are as previously described. Preferred aralkyls comprise alower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude benzyl, 2-phenethyl and naphthalenylmethyl. The bond to theparent moiety is through the alkyl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring systemcomprising about 3 to about 10 carbon atoms, preferably about 5 to about10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7ring atoms. Non-limiting examples of suitable monocyclic cycloalkylsinclude cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.Non-limiting examples of suitable multicyclic cycloalkyls include1-decalinyl, norbornyl, adamantyl and the like.

“Halogen” or “halo” means fluorine, chlorine, bromine, or iodine.

“Heterocyclyl” (or heterocycloalkyl) means a non-aromatic saturatedmonocyclic or multicyclic ring system comprising about 3 to about 10ring atoms, preferably about 5 to about 10 ring atoms, in which one ormore of the atoms in the ring system is an element other than carbon,for example nitrogen, oxygen or sulfur, alone or in combination. Thereare no adjacent oxygen and/or sulfur atoms present in the ring system.Preferred heterocyclyls contain about 5 to about 6 ring atoms. Theprefix aza, oxa or thia before the heterocyclyl root name means that atleast a nitrogen, oxygen or sulfur atom respectively is present as aring atom. The heterocyclyl can be optionally substituted by one or more“ring system substituents” which may be the same or different, and areas defined herein. The nitrogen or sulfur atom of the heterocyclyl canbe optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclylrings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl,tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and thelike.

“Ring system substituent” means a substituent attached to an aromatic ornon-aromatic ring system which, for example, replaces an availablehydrogen on the ring system. Ring system substituents may be the same ordifferent, each being independently selected from the group consistingof alkyl, aryl, heteroaryl, aralkyl, alkylaryl, aralkenyl,heteroaralkyl, alkylheteroaryl, heteroaralkenyl, hydroxy, hydroxyalkyl,alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo, nitro, cyano, carboxy,alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, alkylsulfonyl,arylsulfonyl, heteroarylsulfonyl, alkylsulfinyl, arylsulfinyl,heteroarylsulfinyl, alkylthio, arylthio, heteroarylthio, aralkylthio,heteroaralkylthio, cycloalkyl, cycloalkenyl, heterocyclyl,heterocyclenyl, Y₁Y₂N—, Y₁Y₂N-alkyl-, Y₁Y₂NC(O)— and Y₁Y₂NSO₂—, whereinY, and Y₁ and Y₂ may be the same or different and are independentlyselected from the group consisting of hydrogen, alkyl, aryl, andaralkyl. “Ring system substituent” also means a cyclic ring of 3 to 7ring atoms of which 1-2 may be a heteroatom, attached to an aryl,heteroaryl, heterocyclyl or heterocyclenyl ring by simultaneouslysubstituting two ring hydrogen atoms on said aryl, heteroaryl,heterocyclyl or heterocyclenyl ring. Non-limiting examples include:

and the like.

The term “heteroaralkyl” or “heteroarylalkyl” means aheteroaryl-alkyl-group in which the heteroaryl and alkyl are aspreviously described. Preferred heteroarylalkyls comprise a lower alkylgroup. Non-limiting examples of suitable heteroarylalkyl groups includepyridin-4-ylmethyl, thien-3-ylmethyl and the like. The bond to theparent moiety is through the alkyl.

The term “heterocyclylalkyl” means a heterocyclyl-alkyl-group in whichthe heterocyclyl and alkyl are as previously described. Preferredheterocyclylalkyls comprise a lower alkyl group. Non-limiting examplesof suitable heterocyclylalkyl groups include piperidin-4-ylmethyl,pyrrolidin-3-ylmethyl and the like. The bond to the parent moiety isthrough the alkyl.

The term “substituted” means that one or more hydrogens on thedesignated atom is replaced with a selection from the indicated group,provided that the designated atom's normal valency under the existingcircumstances is not exceeded, and that the substitution results in astable compound. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds. By“stable compound” or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “optionally substituted” means optional substitution with thespecified groups, radicals or moieties.

As a general note to all the Tables that are attached hereto as well asto the Description, Examples and Schemes in this application, anyopen-ended nitrogen atom with unfulfilled valence in the chemicalstructures herein refers to NH, or in the case of a terminal nitrogen,—NH₂. Similarly, any open-ended oxygen atom or carbon atom withunfulfilled valence in the chemical structures herein refers to —OH andany open-ended carbon atom with unfilled valence is appropriately filledwith —H.

When a functional group in a compound is termed “protected”, this meansthat the group is in modified form to preclude undesired side reactionsat the protected site when the compound is subjected to a reaction.Suitable protecting groups will be recognized by those with ordinaryskill in the art as well as by reference to standard textbooks such as,for example, T. W. Greene et al, Protective Groups in organic Synthesis(1991), Wiley, New York.

When any variable (e.g., aryl, heterocycle, R², etc.) occurs more thanone time in any constituent or in Formula I, its definition on eachoccurrence is independent of its definition at every other occurrence.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are alsocontemplated herein. The term “prodrug”, as employed herein, denotes acompound that is a drug precursor which, upon administration to asubject, undergoes chemical conversion by metabolic or chemicalprocesses to yield a compound of Formula III or a salt and/or solvatethereof. A discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, (1987)Edward B. Roche, ed., American Pharmaceutical Association and PergamonPress, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this inventionwith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the present inventioneffective in inhibiting the type 3 17β-hydroxysteroid dehydrogenase andthus producing the desired therapeutic, ameliorative, inhibitory orpreventative effect.

The compounds of formula I form salts which are also within the scope ofthis invention. Reference to a compound of formula I herein isunderstood to include reference to salts thereof, unless otherwiseindicated. The term “salt(s)”, as employed herein, denotes acidic saltsformed with inorganic and/or organic acids, as well as basic saltsformed with inorganic and/or organic bases. In addition, when a compoundof formula I contains both a basic moiety, such as, but not limited to apyridine or imidazole, and an acidic moiety, such as, but not limited toa carboxylic acid, zwitterions (“inner salts”) may be formed and areincluded within the term “salt(s)” as used herein. Pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts arepreferred, although other salts are also useful. Salts of the compoundsof the formula I may be formed, for example, by reacting a compound offormula I with an amount of acid or base, such as an equivalent amount,in a medium such as one in which the salt precipitates or in an aqueousmedium followed by lyophilization. Acids (and bases) which are generallyconsidered suitable for the formation of pharmaceutically useful saltsfrom basic (or acidic) pharmaceutical compounds are discussed, forexample, by S. Berge et al, Journal of Pharmaceutical Sciences (1977)66(1)-1-19; P. Gould, International J. of Pharmaceutics (1986)33201-217; Anderson et al, The Practice of Medicinal Chemistry (1996),Academic Press, New York; in The Orange Book (Food & DrugAdministration, Washington, D.C. on their website); and P. HeinrichStahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts:Properties, Selection, and Use, (2002) Int'l. Union of Pure and AppliedChemistry, pp. 330-331. These disclosures are incorporated herein byreference thereto.

Exemplary acid addition salts include acetates, adipates, alginates,ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates,borates, butyrates, citrates, camphorates, camphorsulfonates,cyclopentanepropionates, digluconates, dodecylsulfates,ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates,hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides,hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates,methanesulfonates, methyl sulfates, 2-naphthalenesulfonates,nicotinates, nitrates, oxalates, pamoates, pectinates, persulfates,3-phenylpropionates, phosphates, picrates, pivalates, propionates,salicylates, succinates, sulfates, sulfonates (such as those mentionedherein), tartarates, thiocyanates, toluenesulfonates (also known astosylates,) undecanoates, and the like.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, aluminum salts, zinc salts, salts withorganic bases (for example, organic amines) such as benzathines,diethylamine, dicyclohexylamines, hydrabamines (formed withN,N-bis(dehydroabietyl)ethylenediamine), N-methyl-D-glucamines,N-methyl-D-glucamides, t-butyl amines, piperazine,phenylcyclohexylamine, choline, tromethamine, and salts with amino acidssuch as arginine, lysine and the like. Basic nitrogen-containing groupsmay be quarternized with agents such as lower alkyl halides (e.g.methyl, ethyl, propyl, and butyl chlorides, bromides and iodides),dialkyl sulfates (e.g. dimethyl, diethyl, dibutyl, and diamyl sulfates),long chain halides (e.g. decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides), aralkyl halides (e.g. benzyl and phenethylbromides), and others.

All such acid salts and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Compounds of formula I, and salts, solvates and prodrugs thereof, mayexist in their tautomeric form (for example, as an amide or iminoether). All such tautomeric forms are contemplated herein as part of thepresent invention.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the present compounds (including those of the salts,solvates and prodrugs of the compounds as well as the salts and solvatesof the prodrugs), such as those which may exist due to asymmetriccarbons on various substituents, including enantiomeric forms (which mayexist even in the absence of asymmetric carbons), rotameric forms,atropisomers, and diastereomeric forms, are contemplated within thescope of this invention, as are positional isomers (such as, forexample, 4-pyridyl and 3-pyridyl). Individual stereoisomers of thecompounds of the invention may, for example, be substantially free ofother isomers, or may be admixed, for example, as racemates or with allother, or other selected, stereoisomers. The chiral centers of thepresent invention can have the S or R configuration as defined by theIUPAC 1974 Recommendations. The use of the terms “salt”, “solvate”“prodrug” and the like, is intended to equally apply to the salt,solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers,positional isomers, racemates or prodrugs of the inventive compounds.

The invention also includes compounds of Formula I in isolated andpurified form.

The compounds according to the invention have pharmacologicalproperties; in particular, the compounds of Formula I can be inhibitorsof type 3 17β-hydroxysteroid dehydrogenase. The novel compounds ofFormula I are expected to be useful in the therapy of proliferativediseases associated with type 3 17β-hydroxysteroid dehydrogenase.

More specifically, the compounds of Formula I can be useful in thetreatment or prevention of androgen or estrogen dependent diseases in apatient in need thereof, which comprises administering to said patient atherapeutically effective amount of at least one compound of formula I.

In another aspect, the invention provides a method of treating orpreventing prostate cancer, and other androgen-dependent neoplasms,benign prostatic hyperplasia, prostatic intraepithelial neoplasia,androgenic alopecia (i.e. pattern baldness in both male and femalepatients), hirsutism, polycystic ovary syndrome and acne in a patient inneed thereof, which comprises administering to said patient, atherapeutically effective amount of at least one compound of formula I.

In another aspect, the invention provides a method of treating orpreventing androgen-dependent diseases in a patient in need thereof,comprising administering (concurrently or sequentially) to said patientan effective amount of at least one compound of formula I in combinationor association with at least one anti-androgenic agent (i.e. agents thatdecrease androgen synthesis or activity).

This invention also provides a method of treating or preventing benignprostatic hyperplasia in a patient in need thereof, comprisingadministering (concurrently or sequentially) to said patient aneffective amount of at least one compound of formula I in combination orassociation with at least one agent useful in the treatment orprevention of benign prostatic hyperplasia.

This invention also provides a method of treating or preventing hairloss in a patient in need thereof, comprising administering(concurrently or sequentially) to said patient an effective amount of atleast one compound of formula I in combination or association with atleast one agent useful in the treatment or prevention of alopecia, e.g.,potassium channel agonists or 5α-reductase inhibitors.

This invention also provides a method of treating or preventing hairloss, comprising administering (concurrently or sequentially) to apatient in need thereof, an effective amount of a compound of formula Iin combination with at least one potassium channel agonist e.g.minoxidil and KC-516, or 5α-reductase inhibitor, e.g., finasteride.

The invention also provides a method of treating or preventingproliferative diseases in a patient in need thereof, especially cancers(tumors), comprising administering (concurrently or sequentially) tosaid patient an effective amount of (1) at least one compound of formulaI in combination or association with (2) an effective amount of at leastone anti-cancer agent i.e., a chemotherapeutic agent, biological agent,and/or surgery, e.g., prostatectomy and/or radiation therapy.

Non-limiting examples of cancers (i.e. tumors) which may be inhibited ortreated include, but are not limited to, lung cancer (e.g., lungadenocarcinoma), pancreatic cancers (e.g., pancreatic carcinoma such as,for example, exocrine pancreatic carcinoma), colon cancers (e.g.,colorectal carcinomas, such as, for example, colon adenocarcinoma andcolon adenoma), renal cancers, myeloid leukemias (for example, acutemyelogenous leukemia (AML), thyroid follicular cancer, myelodysplasticsyndrome (MDS), bladder carcinoma, epidermal carcinoma, melanoma, breastcancer and prostate cancer.

The method of treating proliferative diseases (cancer), according tothis invention, includes a method for treating (inhibiting) the abnormalgrowth of cells, including transformed cells, in a patient in need ofsuch treatment, by administering, concurrently or sequentially, aneffective amount of at least one compound of this invention and aneffective amount of at least one chemotherapeutic agent, biologicalagent, surgery (e.g. prostatectomy) and/or radiation. Abnormal growth ofcells means, for example, cell growth independent of normal regulatorymechanisms (e.g., contact inhibition or apoptosis), including theabnormal growth of: (1) tumor cells (tumors) expressing an activated rasoncogene; (2) tumor cells in which the ras protein is activated as aresult of oncogenic mutation in another gene; and (3) benign andmalignant cells of other proliferative diseases.

In its embodiments, the present invention includes methods for treatingor inhibiting tumor growth in a patient in need of such treatment, byadministering, concurrently or sequentially, (1) an effective amount ofat least one compound of this invention and (2) an effective amount ofat least one antineoplastic/microtubule agent, biological agent, and/orsurgery (e.g. prostatectomy) and/or radiation therapy. Examples oftumors which may be treated include, but are not limited to, epithelialcancers, e.g., prostate cancer, lung cancer (e.g., lung adenocarcinoma),pancreatic cancers (e.g., pancreatic carcinoma such as, for example,exocrine pancreatic carcinoma), breast cancers, renal cancers, coloncancers (e.g., colorectal carcinomas, such as, for example, colonadenocarcinoma and colon adenoma), ovarian cancer, and bladdercarcinoma. Other cancers that can be treated include melanoma, myeloidleukemias (for example, acute myelogenous leukemia), sarcomas, thyroidfollicular cancer, and myelodysplastic syndrome.

As used herein the following terms have the following meanings unlessindicated otherwise:

“Antineoplastic agent” means a chemotherapeutic agent effective againstcancer;

“Concurrently” means (1) simultaneously in time, or (2) at differenttimes during the course of a common treatment schedule; and

“Sequentially” means (1) administration of one component of the method((a) compound of the invention, or (b) antineoplastic agent and/orradiation therapy) followed by administration of the other component;after administration of one component, the second component can beadministered substantially immediately after the first component, or thesecond component can be administered after an effective time periodafter the administration of the first component; the effective timeperiod is the amount of time given for realization of maximum benefitfrom the administration of the first component.

A preferred dosage is about 0.001 to 500 mg/kg of body weight/day of thecompound of Formula I. An especially preferred dosage is about 0.01 to25 mg/kg of body weight/day of a compound of Formula I, or apharmaceutically acceptable salt or solvate of said compound.

Certain useful combination/association agents are described below:

Chemotherapeutic Agents

Classes of compounds that can be used as the chemotherapeutic agent(antineoplastic agent) include: alkylating agents, antimetabolites,natural products and their derivatives, hormones and steroids (includingsynthetic analogs), and synthetics. Non-limiting examples of compoundswithin these classes are:

Alkylating agents (including nitrogen mustards, ethyleniminederivatives, alkyl sulfonates, nitrosoureas and triazenes): Uracilmustard, Chlormethine, Cyclophosphamide (Cytoxan®), Ifosfamide,Melphalan, Chlorambucil, Pipobroman, Triethylenemelamine,Triethylenethiophosphoramine, Busulfan, Carmustine, Lomustine,Streptozocin, Dacarbazine, and Temozolomide.

Antimetabolites (including folic acid antagonists, pyrimidine analogs,purine analogs and adenosine deaminase inhibitors): Methotrexate,5-Fluorouracil, Floxuridine, Cytarabine, 6-Mercaptopurine,6-Thioguanine, Fludarabine phosphate, Pentostatine, and Gemcitabine.

Natural products and their derivatives (including vinca alkaloids,antitumor antibiotics, enzymes, lymphokines and epipodophyllotoxins):Vinblastine, Vincristine, Vindesine, Bleomycin, Dactinomycin,Daunorubicin, Doxorubicin, Epirubicin, Idarubicin, paclitaxel(paclitaxel is commercially available as Taxol® and is described in moredetail below in the subsection entitled “Microtubule Affecting Agents”),Mithramycin, Deoxycoformycin, Mitomycin-C, L-Asparaginase, Interferons-αand β (especially IFN-α), Etoposide, and Teniposide.

Hormonal agents and steroids (including synthetic analogs):17α-Ethinylestradiol, Diethylstilbestrol, Testosterone, Prednisone,Fluoxymesterone, Dromostanolone propionate, Testolactone,Megestrolacetate, Tamoxifen, Methylprednisolone, Methyltestosterone,Prednisolone, Triamcinolone, Chlorotrianisene, Hydroxyprogesterone,Aminoglutethimide, Estramustine, Medroxyprogesteroneacetate, Leuprolide,Flutamide, Toremifene, goserelin and Zoladex.

Synthetics (including inorganic complexes such as platinum coordinationcomplexes): Cisplatin, Carboplatin, Hydroxyurea, Amsacrine,Procarbazine, Mitotane, Mitoxantrone, Levamisole, Navelbene, CPT-11,Anastrazole, Letrazole, Capecitabine, Ralozifine, Droloxifine andHexamethylmelamine.

Non-limiting examples of biological agents useful in the methods of thisinvention include but are not limited to, interferon-α, interferon-β andgene therapy.

Microtubule Affecting Agents

As used herein, a microtubule affecting agent is a compound thatinterferes with cellular mitosis, i.e., having an anti-mitotic effect,by affecting microtubule formation and/or action. Such agents can be,for instance, microtubule stabilizing agents or agents which disruptmicrotubule formation.

Non-limiting examples of microtubule affecting agents useful in theinvention include allocolchicine (NSC 406042), Halichondrin B (NSC609395), colchicine (NSC 757), colchicine derivatives (e.g., NSC 33410),dolastatin 10 (NSC 376128), maytansine (NSC 153858), rhizoxin (NSC332598), paclitaxel (Taxol®, NSC 125973), Taxol® derivatives (e.g.,derivatives (e.g., NSC 608832), thiocolchicine (NSC 361792), tritylcysteine (NSC 83265), vinblastine sulfate (NSC 49842), vincristinesulfate (NSC 67574), epothilone A, epothilone, discodermolide,estramustine, nocodazole, MAP4, and the like.

Particularly preferred agents are compounds with paclitaxel-likeactivity. These include, but are not limited to, paclitaxel andpaclitaxel derivatives (paclitaxel-like compounds) and analogues.Paclitaxel and its derivatives are available commercially. Morespecifically, the term “paclitaxel” as used herein refers to the drugcommercially available as Taxol®.

Examples of such agents include, but are not limited to, inhibitors of5α-reductase type 1 and/or type 2, e.g. finasteride, SKF105,657,LY191,704, LY320,236, dutasteride, flutamide, nicalutamide,bicalutamide, LHRH agonists e.g. leuprolide and zoladex, LHRHantagonists, e.g. abarelix and cetrorelix, inhibitors of17α-hydroxylase/C17-20 lyase, e.g. YM116, CB7630 and liarozole;inhibitors of 17β-hydroxysteroid dehydrogenase type 5 and/or other17β-hydroxysteroid dehydrogenase/17β-oxidoreductase isoenzymes, e.g.EM-1404.

Types of androgen or estrogen dependent diseases include, but are notlimited to, prostate cancer, benign prostatic hyperplasia, prostaticintraepithelial neoplasia, acne, seborrheas, hirsutism, androgenicalopecia, precocious puberty, adrenal hyperplasia, and polycysticovarian syndrome, breast cancer, endometriosis and leiomyoma.

Examples of agents useful in the treatment or prevention of benignprostatic hyperplasia include, but are not limited to, α-1 adrenergicantagonists, e.g. tamsulosin and terazosin.

The pharmacological properties of the compounds of this invention may beconfirmed by a number of pharmacological assays. The exemplifiedpharmacological assays which are described later have been carried outwith the compounds according to the invention and their salts.

This invention is also directed to pharmaceutical compositions whichcomprise at least one compound of Formula I, or a pharmaceuticallyacceptable salt or solvate of said compound and at least onepharmaceutically acceptable carrier.

For preparing pharmaceutical compositions from the compounds describedby this invention, inert, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, dispersible granules, capsules, cachets and suppositories. Thepowders and tablets may be comprised of from about 5 to about 95 percentactive ingredient. Suitable solid carriers are known in the art, e.g.,magnesium carbonate, magnesium stearate, talc, sugar or lactose.Tablets, powders, cachets and capsules can be used as solid dosage formssuitable for oral administration. Examples of pharmaceuticallyacceptable carriers and methods of manufacture for various compositionsmay be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences,18^(th) Edition, (1990), Mack Publishing Co., Easton, Pa.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injection or addition of sweeteners and opacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier, such as an inert compressed gas,e.g. nitrogen.

Also included are solid form preparations that are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions can take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

The compounds of this invention may also be delivered subcutaneously.

Preferably the compound is administered orally.

Preferably, the pharmaceutical preparation is in a unit dosage form. Insuch form, the preparation is subdivided into suitably sized unit dosescontaining appropriate quantities of the active component, e.g., aneffective amount to achieve the desired purpose.

The quantity of active compound in a unit dose of preparation may bevaried or adjusted from about 1 mg to about 100 mg, preferably fromabout 1 mg to about 50 mg, more preferably from about 1 mg to about 25mg, according to the particular application.

The actual dosage employed may be varied depending upon the requirementsof the patient and the severity of the condition being treated.Determination of the proper dosage regimen for a particular situation iswithin the skill of the art. For convenience, the total daily dosage maybe divided and administered in portions during the day as required.

The amount and frequency of administration of the compounds of theinvention and/or the pharmaceutically acceptable salts thereof will beregulated according to the judgment of the attending clinicianconsidering such factors as age, condition and size of the patient aswell as severity of the symptoms being treated. A typical recommendeddaily dosage regimen for oral administration can range from about 1mg/day to about 500 mg/day, preferably 1 mg/day to 200 mg/day, in two tofour divided doses.

The chemotherapeutic agent and/or radiation therapy can be administeredin combination or association with the compounds of the presentinvention according to the dosage and administration schedule listed inthe product information sheet of the approved agents, in the PhysiciansDesk Reference (PDR) as well as therapeutic protocols well known in theart. Table A below gives ranges of dosage and dosage regimens of someexemplary chemotherapeutic agents useful in the methods of the presentinvention. It will be apparent to those skilled in the art that theadministration of the chemotherapeutic agent and/or radiation therapycan be varied depending on the disease being treated and the knowneffects of the chemotherapeutic agent and/or radiation therapy on thatdisease. Also, in accordance with the knowledge of the skilledclinician, the therapeutic protocols (e.g., dosage amounts and times ofadministration) can be varied in view of the observed effects of theadministered chemotherapeutic agents (i.e., antineoplastic agent orradiation) on the patient, and in view of the observed responses of thedisease to the administered therapeutic agents.

TABLE A Exemplary Chemotherapeutic Agents Dosage and Dosage RegimensCisplatin: 50-100 mg/m² every 4 weeks (IV)* Carboplatin: 300-360 mg/m²every 4 weeks (IV) Taxotere: 60-100 mg/m² every 3 weeks (IV)Gemcitabine: 750-1350 mg/m2 every 3 weeks (IV) Taxol: 65-175 mg/m2 every3 weeks (IV) *(IV)-intravenously

Anti-androgenic agents, anti-benign prostatic hyperplasia agents,potassium channel agonists and biological agents can be administered inassociation with the compounds of the present invention according to thedosage and administration schedule listed in the product informationsheet of the approved agents, in the Physicians Desk Reference (PDR) aswell as therapeutic protocols well known in the art. It will be apparentto those skilled in the art that the administration of the agents can bevaried depending on the disease being treated and the known effects ofthe agents on that disease. Also, in accordance with the knowledge ofthe skilled clinician, the therapeutic protocols (e.g., dosage amountsand times of administration) can be varied in view of the observedeffects of the administered agents on the patient, and in view of theobserved responses of the disease to the administered therapeuticagents.

Another aspect of this invention is a kit comprising a therapeuticallyeffective amount of at least one compound of Formula I, or apharmaceutically acceptable salt or solvate of said compound and apharmaceutically acceptable carrier, vehicle or diluent.

Yet another aspect of this invention is a kit comprising an amount of atleast one compound of Formula I, or a pharmaceutically acceptable saltor solvate of said compound and an amount of at least one additionalagent listed above, wherein the amounts of the two or more ingredientsresult in desired therapeutic effect.

The above-described kits may contain the said ingredients in one or morecontainers within said kit.

The invention disclosed herein is exemplified by the followingpreparations and examples which should not be construed to limit thescope of the disclosure. Alternative mechanistic pathways and analogousstructures will be apparent to those skilled in the art.

Where NMR data are presented, ¹H spectra were obtained on either aVarian VXR-200 (200 MHz, ¹H), Varian Gemini-300 (300 MHz) or XL-400 (400MHz) and are reported as ppm down field from Me₄Si with number ofprotons, multiplicities, and coupling constants in Hertz indicatedparenthetically. Where LC/MS data are presented, analyses was performedusing an Applied Biosystems API-100 mass spectrometer and ShimadzuSCL-10A LC column: Altech platinum C18, 3 micron, 33 mm×7 mm ID;gradient flow: 0 min-10% CH₃CN, 5 min-95% CH₃CN, 7 min-95% CH₃CN, 7.5min-10% CH₃CN, 9 min-stop. The retention time and observed parent ionare given.

The following solvents and reagents may be referred to by theirabbreviations in parenthesis:

-   Thin layer chromatography: TLC-   ethyl acetate: AcOEt or EtOAc-   trifluoroacetate: TFA-   triethylamine: TEA-   butoxycarbonyl: n-Boc or Boc-   nuclear magnetic resonance spectroscopy: NMR-   liquid chromatography mass spectrometry: LCMS-   high resolution mass spectrometry: HERMS-   milliliters: mL-   millimoles: mmol-   microliters: μl-   grams; g-   milligrams: mg-   room temperature or rt (ambient): about 25° C.

EXAMPLES

Compounds of formula (l) may be produced by processes known to thoseskilled in the art. Illustrative procedures are shown in therepresentative Schemes, preparations and Examples below. These schemes,preparations and examples should not be construed to limit the scope ofthe disclosure. Alternative mechanistic pathways and analogousstructures may be apparent to those skilled in the art. Some of thecompounds made by these processes are listed in Table 1. As statedearlier, all kinds of isomeric forms of the compounds are considered tobe within the scope of this invention.

The piperidine-piperazine core is added to an appropriate chloride.Deprotection and coupling with N-BOC piperidine acetic acid, followed bydeprotection and acylation, gives the desired product (Scheme 1).

Alternatively, the desired piperidine-piperazine intermediate in Scheme1 can be prepared from an appropriate piperidone (Scheme 2).

Alternatively, the piperidine-piperazine-piperidine core is added to anappropriate chloride to give the desired product, followed bydeprotection and acylation (Scheme 3).

Alternatively, the piperazine-piperidine core is added to an appropriatepiperidone to give the desired product (Scheme 4).

The synthesis of desired chlorides can be accomplished by the reductionof an appropriate ketone (Scheme 5). The resulting alcohol is thenconverted to the requisite chloride under standard conditions. (U.S.Pat. No. 5,719,148).

The substituted piperazines can be prepared through the reduction ofcommercially available diketopiperazines or alternatively from thedesired amino acids (Scheme 6).

The N-BOC or N-acyl piperidine acetic acid can be prepared as describedpreviously through the reduction of 4-pyridine acetic acid (Scheme 7).

Preparative Example 1

To a solution of DCC (43.2 mL, 1.0 M in CH₂Cl₂, 1.0 eq.) in CH₂Cl₂ (200mL) at 0° C. was added N-t-BOC-L-leucine (10 g, 43.2 mmol). To theresulting slurry was added ethyl N-benzylglycinate (8.1 mL, 1.0 eq.)over 15 minutes. The resulting solution was stirred at 0° C. for 2 hoursand room temperature 1 hour, filtered and the concentrated to give acolorless oil (20.7 g, LCMS: MH⁺=407). The intermediate was dissolved inCH₂Cl₂ (150 mL) through which HCl (g) was bubbled for 4 hours. Thesolution was purges with N₂ and concentrated under reduced pressure. Theresidue was neutralized with saturated NaHCO₃ and extracted with EtOAc(3×200 mL). The combined organics were washed with water, dried overNa₂SO₄, filtered and concentrated to give a white solid which was usedwithout further purification (11.3 g, 100% yield). LCMS: MH⁺=261.

Preparative Example 2-4

By essentially the same procedure set forth in Preparative Example 1only substituting the amino acids from Table 1, Column 2, the titlecompounds in Table 1A, Column 3, were prepared:

TABLE 1A Prep. Ex. Column 2 Column 3 DATA 2

LCMS:MH⁺ = 261 3

LCMS:MH⁺ = 261 4

—

Preparative Example 5

To a solution of (S)-3-isopropyl-2,5-piperazinedione (5.0 g, 32 mmol) inTHF (100 mL) at 0° C. was added LAH (137 mL, 1.0 M in THF, 4.3 eq.)dropwise. After the addition was complete, the resulting solution washeated to reflux overnight. The reaction mixture was cooled to roomtemperature and quenched by the slow, sequential addition of water (5.23mL), 1N NaOH (5.23 mL), and water (5.23 mL). The resulting slurry wasdiluted with EtOAc and filtered through a plug of Celite. The residuewas washed with EtOAc (4×100 mL) and the combined organics concentratedunder reduced pressure. The crude product was purified by flashchromatography using a gradient of 5% MeOH, 10% MeOH, 5% (10% NH₄OH) inMeOH, 10% (10% NH₄OH) in MeOH, and 20% (10% NH₄OH) in MeOH in CH₂Cl₂ togive a pale yellow solid (3.03 g, 74% yield). LCMS: MH⁺=129.

Preparative Example 6-11

By essentially the same procedure set forth in Preparative Example 5only substituting the piperazinediones from Table 2, Column 2, the titlecompounds in Table 2, Column 3 were prepared:

TABLE 2 Prep. Ex. Column 2 Column 3 DATA 6

LCMS: MH⁺ = 233 7

LCMS: MH⁺ = 233 8

LCMS: MH⁺ = 233 9

FABMS: MH⁺ =235 10

LCMS: MH⁺ = 143 11

—

Preparative Example 12

To a solution of N-Boc-4-piperidineacid acid (prepared as described inU.S. Pat. No. 5,874,442; 35.0 g, 144 mmol) and TEA (20.0 mL, 1.0 eq.) intoluene (100 mL) at 0° C. was added trimethylacetyl chloride (17.7 mL,1.0 eq.). The resulting slurry was stirred at 0° C. 1.5 hours beforeadding the title compound from Preparative Example 8 (33.5 g, 151 mmol,1.05 eq.) in toluene (100 mL) and the resulting solution was warmed toroom temperature and stirred overnight. The reaction mixture wasneutralized by the addition of 1N NaOH and extracted with EtOAc. Thecombined organics were dried over Na₂SO₄, filtered, and concentrated.The crude product was purified by flash chromatography using a 50:50EtOAc:Hexane solution as eluent (34.4 g, 51% yield). LCMS: MH⁺=458.

Preparative Example 13

To a solution of the title compound from Preparative Example 12 (34.0 g,74.3 mmol) in absolute EtOH (600 mL) was added 10% Pd—C (35.0 g, wet,50%) and NH₄HCO₂ (94 g, 10 eq.). The reaction mixture was heated toreflux for 3 hours, cooled to room temperature, filtered through a plugof Celite and concentrated under reduced pressure. The residue wasdiluted with EtOAc and washed sequentially with H₂O, 1N NaOH, H₂O, andbrine. The organics were dried over Na₂SO4, filtered, and concentratedin vacuo. The crude product was purified by flash chromatography using a5% (10% NH₄OH in MeOH) in CH₂Cl₂ to 10% (10% NH₄OH in MeOH in CH₂Cl₂ aseluent (20 g, 74% yield). LCMS: MH⁺=368.

Preparative Example 14

To a solution of the title compound from Preparative Example 7 (8.2 g,31.5 mmol) in CH₂Cl₂ (300 mL) was added (BOC)₂O (7.5 g, 1.02 eq.). Theresulting solution was stirred at room temperature overnight. Thereaction quenched by the addition of saturated NaHCO₃ and separated. Theorganic layer was washed with brine, dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The crude product was purified byflash chromatography using a 10% EtOAc in hexanes solution as eluent(10.6 g, 99% yield). LCMS: MH⁺=333.

Preparative Example 15

By essentially the same procedure set forth in Preparative Example 14,only substituting the title compound from Preparative Example 9 in Table3, Column 2, the title compounds in Table 3, Column 3 was prepared:

TABLE 3 Prep. Ex. Column 2 Column 3 DATA 15

LCMS:MH⁺ = 335

Preparative Example 16

A solution of the title compound from Preparative Example 14 (10.4 g,31.3 mmol) and 10% Pd/C (1.95 g) in EtOH (130 mL) was hydrogenated on aParr apparatus at 50 psi overnight The reaction mixture was filteredthrough Celite and the filtrate concentrated in vacuo to give theproduct as a colorless oil (6.93 g, 91% yield) which was used withoutfurther purification. LCMS: MH⁺=243.

Preparative Examples 17-20

By essentially the same procedure set forth in Preparative Example 16only substituting compounds from Table 4, Column 2, the title compoundsin Table 4, Column 3 were prepared:

TABLE 4 Prep. Ex. Column 2 Column 3 DATA 17

LCMS:MH⁺ = 245 18

LCMS:MH⁺ = 368 19

LCMS:MH⁺ = 356 20

LCMS:MH⁺ = 368

Preparative Example 21

4,4′-Bipiperidine (17.5 g, 72.55 mmol) was dissolved in H₂O (70 mL),treated with 5 N NaOH to pH 8-9, and diluted to 400 mL with EtOH. To thestirred mixture at room temperature was added di-t-butyl dicarbonate(16.8 g, 76.96 mmol) in 200 mL of EtOH in one portion. The reactionmixture was treated with 5 N NaOH periodically to pH 8-9. After 5 hours,the mixture was concentrated. The residue was dissolved in 500 mL of 1:1H₂O:Et₂O and the PH was adjusted to 12 with 5 N NaOH. The aqueous phasewas extracted with Et₂O and combined organic phase was washed withbrine, 5% aq. citric acid. The citric acid washing solution was adjustedto PH 12-13 with 5 N NaOH and extracted with 250 mL of Et₂O 3 times. Thecombined Et₂O was washed with brine, dried over MgSO₄ and concentratedunder vacuum to give the desired product (8.61 g, 44.2% yield). FABMS:MH⁺=269.

Preparative Example 22

Step A:

1-Benzyl-4-piperidone (4.00 g, 21.13 mmol), t-BOC-piperazine (3.94 g,21.14 mmol) and Ti(OiPr)₄ (7.5 g, 26.38 mmol) were stirred at r.t. underN₂ in dry CH₂Cl₂ (8 mL) overnight. To the reaction absolute EtOH (60 mL)was added, followed by NaCNBH₃ (1.32 g, 21 mmol). The mixture wasstirred at r.t. for 6 hrs, diluted with 10 mL of EtOAc, added 20 gfilter celite, 100 mL of ice-cold H₂O and stirred for 1 hr. The mixturewas then filtered and cake was washed with CH₂Cl₂. The combined CH₂Cl₂was washed with brine, dried over Na₂SO₄, filtered, and concentrated.The crude product was purified by flash chromatography using a 3-5% MeOHin CH₂Cl₂ to yield a colorless solid (2.45 g, 32% yield). FABMS:MH⁺=360.

Step B:

To a solution of the title compound from Preparative Example 22, Step A(1.29 g, 3.59 mmol) and Pd/C (0.4 g, 10%) in MeOH (25 mL) was added 96%HCOOH (1 mL, 5.0 eq.). The resulting mixture was stirred at r.m.overnight and filtered through a celite pad and washed with MeOH. Thecombined filtrate was concentrated to syrup, 5 mL of water was added toit and adjusted to PH 12 with 15% NaOH. The mixture was extracted withEtOAc (30 mL) twice and EtOH:CH₂Cl₂ 1:4 (50 mL) twice. The combinedorganics were dried over Na₂SO₄, filtered, and concentrated to yield acolorless solid (0.91 g, 94% yield). FABMS: MH⁺=270.

Preparative Example 23

Step A:

To a solution of the title compound from Preparative Example 21 (2.66 g,9.93 mmol) in 40 mL of 50% aq. MeOH was added Na₂CO₃ (2.1 g). followedby dropwise addition of CbzCl (1.7 mL, 11.9 mmol). The resulting mixturewas stirred at 0° C. for 1 hr. and r.t. for 24 hrs. 100 mL of distilledwater was added to the reaction and the mixture was extracted with EtOAc(100 mL) twice. The crude product was purified by flash chromatographyusing a 1% MeOH in CH₂Cl₂ to yield a white solid (2.97 g). FABMS:MH⁺=403.

Step B:

To a solution of the title compound from Preparative Example 23, STEP A(1.3 g, 3.23 mmol) in CH₂Cl₂ (40 mL) at 0° C. was added dropwise TFA (31mL) over 10 min. The resulting mixture was stirred at 0-5° C. for 2 hrs,then concentrated under reduced pressure The residue was dissolved inCH₂Cl₂/H₂O, adjusted pH to 12 by the addition of 5N NaOH, separated,extracted aqueous layer with CH₂Cl₂. The combined organics were driedover Na₂SO₄, filtered and concentrated to give a white solid (0.885 g)which was used without further purification. FABMS: MH⁺=303.

Step C:

A solution of the title compound from Preparative Example 23, STEP B,N-Boc-4-piperidineacid acid (prepared as described in U.S. Pat. No.5,874,442; 0.1.24 g), DEC (0.86 g), HOBt (0.6 g), and NMM (0.75 mL) inanhydrous DMF (20 mL) was stirred at room temperature for 24 hrs. Thereaction was quenched by the addition of 1N NaOH and extracted withCH₂Cl₂. The combined organics were dried over Na₂SO₄, filtered, andconcentrated. The crude product was purified by flash chromatographyusing a 10% acetone in hexane solution as eluent to give the desiredproduct (1.75 g, 98% yield); FABMS: MH⁺=528.

Step D:

A suspension of the title compound from Preparative Example 23, STEP C(1.18 g, 2.24 mmol) and 10% Pd/C (0.2 g) in MeOH (25 mL) washydrogenated at room temperature for 2 hr. Catalysts were filtered offthrough celite. The filtrate was concentrated to give a syrup, which wasused without further purification. FABMS: MH⁺=394.

Preparative Example 24

Step A:

To a solution of piperidine-4-acetic acid (10.0 g, 70.0 mmol) was inEtOH (100 mL) was added concentrated HCl (2.68 mL, 2.2 eq.). Theresulting solution was heated at reflux for 12 hours. The reactionmixture was concentrated under reduced pressure and used without furtherpurification (10 g, 84% yield).

Step B:

To a solution of the title compound from Preparative Example 24, Step A(2.0 g, 9.6 mmol) in CH₂Cl₂ (30 mL) at 0° C. was added TMSNCO (6.3 mL,5.0 eq.) followed by TEA (2.0 mL, 1.5 eq.). The resulting solution wasstirred at 0° C. for 3 hours and quenched by the addition water anddiluted with saturated NaHCO₃. The mixture was extracted with CH₂Cl₂ andthe combined organics dried over Na₂SO₄, filtered, and concentrated. Thecrude product was purified by flash chromatography using an 8% (10%NH₄OH in MeOH) in CH₂Cl₂ as eluent (1.2 g, 60% yield). FABMS: MH⁺=215.

Step C:

A solution of the title compound from Preparative Example 24, Step B(1.23 g, 5.7 mmol) and LiOH (0.33 g, 2.4 eq.) in CH₂Cl₂ (29 mL), EtOH(29 mL) and water (14 mL) was heated at reflux 3 hours. The resultingsolution was cooled to room temperature, neutralized by the addition of1N HCl (16.1 mL, 2.98 eq.) and concentrated under reduced pressure. Thereaction product was further dried by the azeotropic removal of waterwith toluene to yield an off-white gum (1.1 g, quantitative yield).FABMS: MH⁺=187.

Preparative Example 25

By essentially the same procedure set forth in Preparative Example 23,STEP C, only substituting the title compound from Preparative Example 24and piperidine-piperidine compound (prepared according to knownprocedures) gave the title compound.

Preparative Example 26

By essentially the same procedure set forth in Preparative Example 23,STEP B only substituting the title compound from Preparative Example 25,gave the title compound. LCMS: MH⁺=366.

Preparative Example 27

A solution of the piperazine-anhydride compound (prepared according toknown procedures, 0.2 g, 0.78 mmol) in EtOH (5 mL) was heated at refluxuntil the starting material was gone. The resulting solution was cooledto room temperature and concentrated under reduced pressure to yield thedesired product.

Preparative Example 28

By essentially the same procedure set forth in Preparative Example 23,STEP C, only substituting the title compound from Preparative Example27, gave the title compound, mp=70-76° C. FABMS: MH⁺=427.

Preparative Example 29

By essentially the same procedure set forth in Preparative Example 23,STEP B, only substituting the title compound from Preparative Example28, gave the title compound. LCMS: MH⁺=327.

Preparative Examples 30-37

The synthesis of desired chlorides in Table 5, Column 2 is described incorresponding patents and patent applications (see, for example, U.S.Pat. No. 5,719,148).

TABLE 5 Prep. Ex. Chloride Reference 30

31

32

33

34

35

36

37

Preparative Example 38

To a solution of the title compound from Example 21 (0.47 g, 1.75 mmol,1.135 eq.) and Et₃N (0.24 mL) in anhydrous CH₂Cl₂ (20 mL) was added thefreshly prepared compound from Preparative Example 32 (1.54 mmol) inanhydrous CH₂Cl₂ (5 mL). The resulting mixture was stirred at r.t. for16 hrs and the solvent was evaporated under vacuum. The residue waspurified by flash chromatography using 5%, 10% and 15% EtOAc in Hexaneas eluent to give a white puff solid (0.8 g, 90% yield). FABMS: MH⁺=574.

Preparative Example 39

By essentially the same procedure set forth in Preparative Example 38,only substituting the title compound from Preparative Example 22 (0.475g, 1.76 mmol), using 3% MeOH/ CH₂Cl₂ as eluent, gave the title compound(0.65 g, 73% yield). FABMS: MH⁺=575.

Preparative Example 40-46

By essentially the same procedure set forth in Preparative Example 38 orPreparative Example 39, only substituting the chlorides from Table 6,Column 2, the title compounds in Table 6, Column 3 were prepared:

TABLE 6 Prep. Ex. Column 2 Column 3 DATA 40

FABMS:MH⁺ = 652 41

FABMS:MH⁺ =496. 42

FABMS:MH⁺ =574. 43

FABMS:MH⁺ =652. 44

FABMS:MH⁺ =530. 45

FABMS:MH⁺ =652. 46

FABMS:MH⁺ =606.

Preparative Example 47

To a solution of 4-piperidone (0.2 g, 1.3 mmol) in DMF (5 mL) was addedEt₃N (0.95 mL, 6.5 mmol, 5 eq.) and stirred for 0.5 hr. To the reactionmixture was added the title compound from Preparative Example 30 (0.67g, 1.95 mmol, 1.5 eq.) and stirred overnight. The reaction was quenchedby the addition of saturated NaHCO₃ and extracted with CH₂Cl₂. Thecombined organics were dried over Na₂SO₄, filtered and concentrated. Thecrude product was purified by flash chromatography using a 3% EtOAc inCH₂Cl₂ solution as eluent to yield the desired product (0.4 g, 73%yield). FABMS: MH⁺=405.

Preparative Example 48

To a solution of the title compound from Preparative Example 19 (0.15 g,0.617 mmol) and the title compound from Preparative Example 47 (0.254 g,0.629 mmol, 1.02 eq.) in dry CH₂Cl₂ (5 mL) was added NaBH(OAc)₃, aceticacid (0.05 mL) and stirred for 3 days. The reaction was basified by theaddition of 50% NaOH and extracted with CH₂Cl₂. The combined organicswere washed with brine, dried over Na₂SO₄, filtered and concentrated.The crude product was purified by flash chromatography using a 3%MeOH/CH₂Cl₂ as eluent to yield the white solid (0.163 g, 42% yield).mp=83-87° C., FABMS: MH⁺=633.

Preparative Examples 49-52

By essentially the same procedure set forth in Preparative Example 8only substituting the piperazine from Column 2 of Table 7, the titlecompounds in Column 3, Table 7 were prepared.

TABLE 7 Prep. Ex. Column 2 Column 3 DATA 49

FABMS:MH⁺ =631. 50

LCMS:MH⁺ =517. 51

LCMS:MH⁺ =531. 52

LCMS:MH⁺ =565.

Preparative Example 53

To a solution of the title compound from Preparative Example 38 (0.72 g,1.25 mmol) in CH₂Cl₂ (20 mL) at 0° C. was added dropwise TFA (11 mL).The resulting mixture was stirred at 0-5° C. for 2 hrs, thenconcentrated under reduced pressure. The residue was dissolved inCH₂Cl₂/H₂O, adjusted PH to 13 by the addition of 1N NaOH, separated,extracted aqueous layer with CH₂Cl₂. The combined organics were driedover Na₂SO₄, filtered and concentrated to give a white solid (0.59 g,99% yield) which was used without further purification. FABMS: MH⁺=474.

Preparative Example 54-63

By essentially the same procedure set forth in Preparative Example 53only substituting the compounds from Table 8, Column 2, the titlecompounds in Table 8, Column 3 were prepared:

TABLE 8 Prep. Ex. Column 2 Column 3 DATA 54

FABMS:MH⁺ = 475 55

FABMS:MH⁺ = 552 56

FABMS:MH⁺ =396. 57

FABMS:MH⁺ =474. 58

FABMS:MH⁺ =552. 59

FABMS:MH⁺ =430. 60

FABMS:MH⁺ =552. 61

FABMS:MH⁺ =506. 62

LCMS:MH⁺ =533. 63

FABMS:MH⁺ =531.

Preparative Examples 64 and 65

The title compounds were prepared through the separation ofdiastereomers of the title compound from Preparative Example 53 bypreparative HPLC with a CHIRALPAK AD column using a 85:15 Hexane:IPAwith 0.2% DEA as eluent: Preparative Example 64 [first eluting isomer(+)]: FABMS: MH⁺=474. Preparative Example 65 [first eluting isomer (−)]:FABMS: MH⁺=474.

Preparative Example 66

A solution of the title compound from Preparative Example 53 (0.26 g,0.55 mmol), N-Boc-4-piperidineacid acid (prepared as described in U.S.Pat. No. 5,874,442; 0.2 g, 0.822 mmol), DEC (0.14 g, 0.73 mmol), HOBt(0.096 g, 0.71 mmol), and NMM (0.1 mL, 0.91 mmol) in anhydrous DMF (8.0mL) was stirred at room temperature for 24 hrs. The reaction wasquenched by the addition of 1N NaOH and extracted with CH₂Cl₂. Thecombined organics were dried over Na₂SO₄, filtered, and concentrated.The crude product was purified by flash chromatography using a 2% MeOHin CH₂Cl₂ solution as eluent to give a yellow puff solid (1.38 g, 99%yield). FABMS: MH⁺=699.

Preparative Example 67-81

By essentially the same procedure set forth in Preparative Example 66only substituting the compounds from Table 9, Column 2, the titlecompounds in Table 9, Column 3 were prepared:

TABLE 9 Prep. Ex. Column 2 Column 3 DATA 67

FABMS:MH⁺ =699. 68

FABMS:MH⁺ =699. 69

FABMS:MH⁺ =700 70

FABMS:MH⁺ =777. 71

FABMS:MH⁺ =621. 72

FABMS:MH⁺ =699. 73

FABMS:MH⁺ =777. 74

FABMS:MH⁺ =655. 75

FABMS:MH⁺ =777. 76

LCMH⁺:MH⁺ =731. 77

LCMS:MH⁺ =758. 78

LCMS:MH⁺ =756. 79

LCMS:MH⁺ =742. 80

LCMS:MH⁺ =756. 81

LCMS:MH⁺ =790.

Preparative Example 82

Alternatively, by essentially the same procedure set forth inPreparative Example 38, only substituting the title compound fromPreparative Example 36 and the title compound from Preparative Example25, gave the title compound of this Example (1.09 g, 62% yield). FABMS:MH⁺=777.

Example 100

By essentially the same procedure set forth in Preparative Example 82only substituting the compounds from Table 10, Column 2, the compoundsfrom Table 10, Column 3, the title compounds in Table 10, Column 4 wereprepared:

TABLE 10 Ex. Column 2 Column 3 Column 4 DATA 100

LCMS:MH⁺ =670;m.p. =108-130° C.

Preparative Example 83 and 84

The title compounds were prepared through the separation ofdiastereomers of the title compound from Preparative Example 82 bypreparative HPLC with a CHIRALPAK AD column using a 85:15 Hexanes:IPAwith 0.2% DEA as eluent: Preparative Example 83 [first eluting isomer(+)]: FABMS: MH⁺=777. Preparative Example 84 [second eluting isomer(−)]: FABMS: MH⁺=777.

Preparative Examples 85 and 86

The title compounds were prepared through the separation ofdiastereomers of the title compound from Preparative Example 79 bypreparative HPLC with a CHIRALPAK AD column using a 85:15 Hexane:IPAwith 0.2% DEA as eluent: Preparative Example 85 (first eluting isomer):LCMH⁺: MH⁺=742. Preparative Example 86 (second eluting isomer): LCMH⁺:MH⁺=742.

Preparative Example 87 and 88

The title compounds were prepared through the separation ofdiastereomers of the title compound from Preparative Example 80 bypreparative HPLC with a CHIRALPAK AD column using a 85:15 Hexane:IPAwith 0.2% DEA as eluent: Preparative Example 87 (first eluting isomer):LCMH⁺: MH⁺=756. Preparative Example 88 (second eluting isomer): LCMH⁺:MH⁺=756.

Preparative Example 89

To a solution of the title compound from Preparative Example 76 (0.261g, 0.36 mmol) in toluene (15 mL) and MeOH (5 mL) was added PPh₃ (0.037g, 0.14 mmol), DBU (0.061 g, 0.40 mmol) and PdCl₂ (0.005 g, 0.028 mmol).The resulting mixture was transferred to a CO reactor and heated to 80°C. at 100 psi of CO for 3.5 hrs. The reaction was cooled to r.t.,stirred at r.t. for 2 days and concentrated under reduced pressure. Theresidue was dissolved in 150 mL of EtOAc, washed with H₂O, dried overNa₂SO₄, filtered and concentrated. The crude product was purified byflash chromatography using a 30% EtOAc in CH₂Cl₂ solution as eluent togive the desired product.

Preparative Example 90

By essentially the same procedure set forth in Preparative Example 23,STEP B, only substituting the title compound from Preparative Example 68(0.46 g, 0.66 mmol) gave the title compound (0.39 g, 99% yield). FABMS:MH⁺=599.

Preparative Example 91-112

By essentially the same procedure set forth in Preparative Example 90only substituting the BOC-compounds from Table 11, Column 2, the titlecompounds in Table 11, Column 3 were prepared:

TABLE 11 Prep. Ex. Column 2 Column 3 DATA 91

FABMS:MH⁺ =599. 92

FABMS:MH⁺ =599. 93

FABMS:MH⁺ =600. 94

FABMS:MH⁺ =677. 95

FABMS:MH⁺ =521. 96

FABMS:MH⁺ =599. 97

FABMS:MH⁺ =677. 98

FABMS:MH⁺ =677. 99

FABMS:MH⁺ =677. 100

FABMS:MH⁺ =555. 101

FABMS:MH⁺ =677. 102

LCMH⁺MH⁺ =611. 103

LCMH⁺MH⁺ =597. 104

LCMH⁺MH⁺ =658. 105

LCMH⁺MH⁺ =656. 106

LCMH⁺MH⁺ =642. 107

LCMH⁺MH⁺ =656. 108

LCMH⁺MH⁺ =642. 109

LCMH⁺MH⁺ =642. 110

LCMH⁺MH⁺ =656. 111

LCMH⁺MH⁺ =656. 112

LCMH⁺ =690

Example 200

To a solution of the title compound from Preparative Example 90 (0.14 g,0.233 mmol) in CH₂Cl₂ (4 mL) at r.t. under N₂ was added 85% TMSNCO (0.25mL, 1.89 mmol). The reaction mixture was stirred at r.t. overnight. Thereaction was quenched by the addition of saturated NaHCO₃ and extractedwith CH2Cl₂. The combined organics were dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by flash chromatographyusing a 5% (10% NH₄OH in MeOH) in CH₂Cl₂ solution as eluent to yield thewhite amorphous powder (0.105 g, 70% yield). FABMS: MH⁺=642.

Examples 300-2400

By essentially the same procedure set forth in Example 200 onlysubstituting the compounds from Table12, Column 2, the title compoundsin Table 12, Column 3 were prepared:

TABLE 12 Prep. Ex. Column 2 Column 3 DATA 300

FABMS:MH⁺ =642. 400

FABMS:MH⁺ =642. 500

FABMS:MH⁺ =643. 600

FABMS:MH⁺ =595. 700

FABMS:MH⁺ =720. 800

FABMS:MH⁺ =564. 900

FABMS:MH⁺ =642;mp = . 1000

FABMS:MH⁺ =720. 1100

FABMS:MH⁺ =720. 1200

FABMS:MH⁺ =720. 1300

FABMS:MH⁺ =598;m.p. =131-140° C. 1400

FABMS:MH⁺ =700; 1500

LCMS:MH⁺ =654;m.p. =136.8-139.2° C. 1600

LCMS:MH⁺ =701.m.p. =93-114° C. 1700

LCMS:MH⁺ =699;m.p. =97.2-97.9° C. 1800

LCMS:MH⁺ =685;m.p. =93-120° C. 1900

LCMS:MH⁺ =699;m.p. =59-94° C. 2000

LCMS:MH⁺ =685;m.p. =115-126° C. 2100

LCMS:MH⁺ =685;m.p. =84-110° C. 2200

LCMS:MH⁺ =699;m.p. =120-124° C. 2300

LCMS:MH⁺ =699;m.p. =106.4-111° C. 2400

LCMS:MH⁺ =733;m.p. =98-107° C.

Example 2500

By essentially the same procedure set forth in Preparative Example 48only substituting the piperazine from the title compound of PreparativeExample 29, gave the desired product. m.p.=91-127° C.; LCMS: MH⁺=715.

Example 2600

To a solution of the title compound from Example 1500 (0.05 g, 0.08mmol) in EtOH (1 mL) and H₂O (1 mL) was added LiOH.2H₂O and stirred atr.t. overnight. A few drops of 1 M HCl was added to the reaction untilit was slightly acidic. The reaction was concentrated and purified byflash chromatography using a 50% MeOH/0.1% HOAc/H₂O as eluent to yieldsolid (0.042 g, 83% yield). m.p.=131.9-134.8° C. FABMS: MH⁺=640.

Example 2700

To a solution of the title compound from Preparative Example 100 (0.096g, 0.153 mmol) in CH₂Cl₂ (5 mL) at 0° C. was added TEA (0.039 mL, 5 eq.)and AcCl (0.016 mL, 1.2 eq.). The reaction mixture was stirred at r.t.for 2 hrs. The reaction was quenched by the addition of saturated NaHCO₃and extracted with CH₂Cl₂. The combined organics were dried over Na₂SO₄,filtered and concentrated. The crude product was purified by flashchromatography using a 7% MeOH in CH₂Cl₂ solution as eluent to yield awhite solid (0.061 g, 66% yield). m.p.=94-101° C.; FABMS: MH⁺=597.

Examples 2800-3400

By essentially the same procedure set forth in Example only substitutingthe compound from Table 13, Column, the title compounds shown in Table13, column 3 were prepared:

TABLE 13 Prep. Ex. Column 2 Column 3 DATA 2800

LCMS:MH⁺ =516;m.p. =92-97° C. 2900

LCMS:MH⁺ =698;m.p. =122.3-123.5° C. 3000

LCMS:MH⁺ =684;m.p. =90-104° C. 3100

LCMS:MH⁺ =698;m.p. =75-87° C. 3200

LCMS:MH⁺ =698m.p. =113-123° C. 3300

LCMS:MH⁺ =698;m.p. =120-130° C. 3400

LCMS:MH⁺ =732;m.p. =115-123° C.

Examples 3500 and 3600

The title compounds were prepared through the separation ofdiastereomers of the title compound from Example 3000 by preparativeHPLC with a CHIRALPAK AD column using a 85:15 Hexane:IPA with 0.2% DEAas eluent:

-   Preparative Example 3500 (first eluting isomer): FABMS: MH⁺=684;    m.p.=89-111° C.-   Preparative Example 3600 (second eluting isomer): FABMS: MH⁺=684;    m.p.=93-105° C.

The compounds of the invention can be useful as inhibitors of type 317β-hydroxysteroid dehydrogenase. This utility was demonstrated by thefollowing assay.

Biological Data

17β-Hydroxysteroid Dehydrogenase Inhibition Data Methods:

To prepare human recombinant type 3 17β-hydroxysteroid dehydrogenaseenzyme, HEK-293 cells stably transfected with human 17β-HSD type 3 werecultured to confluency and harvested for enzyme. The cells weresuspended in isolation buffer (20 mM KH₂PO₄, 1 mM EDTA, 0.25 M Sucrose,1 mM PMSF, 5 □g/ml pepstatin A, 5 μg/ml antipain and 5 □g/ml leupeptin)to a concentration between 5.0×10⁶ and 1.0×10⁷ cells/ml. The cells weresonicated on ice using a micro-ultrasonic cell disrupter at an outputsetting of No. 40 for four 10 second bursts. The broken cells were thencentrifuged at 100,000 ×g for 60 min at 4° C., and the resulting pelletwas resuspended, aliquoted into microfuge tubes, and stored at −80° C.

To measure conversion of ¹⁴C-androstenedione to ¹⁴C-testosterone,reaction buffer (12.5 mM KH₂PO₄, 1 mM EDTA), NADPH cofactor (1 mMfinal), test compound, 17β-HSD3 enzyme (30 μg protein) and¹⁴C-androstenedione substrate (100 nM; 2.7 nCi/tube) were added to13×100 borosilicate glass tubes to a total volume of 0.5 mL/tube. Thetubes were placed in a prewarmed 37° C. water bath for 30 minutes. Thereaction was then stopped by adding 1 ml of ethyl ether. The tubes werecentrifuged for 20 minutes at 3000 rpm at 4° C. in a table topcentrifuge and then snap frozen in a dry ice-methanol bath. The etherlayer was decanted into another glass tube, and then evaporated todryness using compressed nitrogen gas. The samples were resuspended inchloroform (20 mL) and spotted onto silica G60 thin layer chromatographyplates. ¹⁴C-Androstenedione substrate and ¹⁴C-testosterone product wereseparated by placing the plates in chloroform:ethyl acetate (3:1). Theplates were dried, exposed overnight, scanned and quantitated on a FUJIFLA2000 phosphorimager.

Compounds of this invention exhibited a range of 17β-hydroxysteroiddehydrogenase Type 3 binding activity from about 0.025 nM to about >100nM. Several compounds of this invention have a binding activity in therange of about 0.025 nM to 10 nM.

While the present invention has been described with in conjunction withthe specific embodiments set forth above, many alternatives,modifications and other variations thereof will be apparent to those ofordinary skill in the art. All such alternatives, modifications andvariations are intended to fall within the spirit and scope of thepresent invention.

1. A method at inhibiting type 3 17β-hydroxysteroid dehydrogenase,comprising administering a therapeutically effective amount of at leastone compound represented by the structural formula:

wherein: X is CH; Y is N; G is C(═O); R is selected from the groupconsisting of alkyl, —OR⁴, aryl, heteroaryl, heteroaryloxy,heterocyclyl, heterocyclyloxy, cycloalkyl, cycloalklyloxy, —N(R⁴)₂ wherethe two R⁴ moieties can be the same or different, —(CH₂)_(n)-aryl,—(CH₂)_(n)-heteroaryl, —(CH₂)_(n)-heterocyclyl and—(CH₂)_(n)-cycloalkyl, wherein each of said alkyl, aryl, heteroaryl, andcycloalkyl can be unsubstituted or optionally independently substitutedwith one or more moieties which can be the same or different, eachmoiety being independently selected from the group consisting of alkyl,alkyl, aryl, heteroaryl, —OR⁴, heterocyclyl, heterocyclyloxy,cycloalkyl, cycloalklyloxy, —N(R⁴)₂ where the two R⁴ groups can be thesame or different, —C(O)R⁴, and —C(O)N(R⁴)₂ where the two R⁴ moietiescan be the same or different; one of a, b, c and d in ring I representsN or N⁺O⁻, and the remaining a, b, c and d positions represent C(R¹) orC(R²); or each of a, b, c, and d are independently selected from C(R¹)or C(R²); R¹ and R² can be the same or different, each beingindependently selected from the group consisting of: H, halo, —CF₃,—OR⁴, —C(O)R⁴, —OCF³, —SR⁴, —S(O)_(n)R⁵, benzotriazol-1-yloxy,tetrazol-5-ylthio, alkynyl, alkenyl wherein said alkenyl can beunsubstituted or optionally substituted with halo, —OR⁴ or —C(O)OR⁴,alkyl wherein said alkyl can be unsubstituted or optionally substitutedwith halo, —OR⁴ or —C(O)OR⁴, —N(R⁴)₂ where the two R⁴ moieties can bethe same or different, —NO₂, —OC(O)R⁵, —C(O)OR⁴, —CN, −N(R⁴)C(O)OR⁴,—SR⁵C(O)OR⁴, and —SR⁵N(R⁴)₂ (provided that R⁵ in —SR⁵N(R⁴)₂ is not—CH₂—) wherein each R⁴ is independently selected; the dotted linebetween carbon atoms 5 and 6 represents an optional bond, such that whena double bond is present, A and B can be the same or different, eachbeing independently selected from the group consisting of —R⁴, halo,—OR⁴, —C(O)OR⁴, —OC(O)OR⁴ or —OC(O)R⁴, and when no double bond ispresent between carbon atoms 5 and 6, A and B can be the same ordifferent, each being independently selected from the group consistingof (H₂), —(OR⁵)₂, (H and halo), (dihalo), (H and R⁵), (R⁵)₂, (H and—OC(O)R⁴), (H and —OR⁴), (═O), and (H, (═NOR⁴) or (—O—(CH₂)_(p)—O—)wherein p is 2, 3 or 4); R³ is selected from the group consisting of H,alkyl, alkoxy and alkoxyalkyl; R⁴ is selected from the group consistingof H, alkyl, aryl and aralkyl; R⁵ is alkyl or aryl; R⁶ is H or alkyl; nis a number from 1-4; and q is a number from 1-8; to a patient in needof such inhibition.
 2. A method of inhibiting type 3 17β-hydroxysteroiddehydrogenase, comprising administering a therapeutically effectiveamount of at least one compound represented by the structural formula:

or a pharmaceutically acceptable salt thereof to a patient in need ofsuch inhibition.